Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Measuring hemoglobin in plasma by reaction with tetramethylbenzidine

S S Levinson, J Goldman

    Clinical Chemistry
    |March 1, 1982
    PubMed
    Summary
    This summary is machine-generated.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Results from a phase Ib study of telisotuzumab vedotin in combination with osimertinib in patients with c-Met protein-overexpressing, EGFR-mutated locally advanced/metastatic non-small-cell lung cancer (NSCLC) after progression on prior osimertinib.

    Annals of oncology : official journal of the European Society for Medical Oncology·2025
    Same author

    Thrombogenicity of biodegradable metals.

    Bioactive materials·2024
    Same author

    Long-term survival with first-line nivolumab plus ipilimumab in patients with advanced non-small-cell lung cancer: a pooled analysis.

    Annals of oncology : official journal of the European Society for Medical Oncology·2022
    Same author

    Absence of damaging effects of stem cell donation in unrelated donors assessed by FISH and gene variance screening.

    Bone marrow transplantation·2020
    Same author

    Comparison of sporadic and familial behavioral variant frontotemporal dementia (FTD) in a North American cohort.

    Alzheimer's & dementia : the journal of the Alzheimer's Association·2020
    Same author

    Porous biodegradable EW62 medical implants resist tumor cell growth.

    Materials science & engineering. C, Materials for biological applications·2016

    This study reveals how albumin affects hemoglobin assays using tetramethylbenzidine. Optimizing incubation conditions allows for accurate hemoglobin measurement across various analytical instruments.

    Area of Science:

    • Clinical Chemistry
    • Biochemistry

    Background:

    • Hemoglobin assays are crucial for diagnosing anemia and monitoring treatment.
    • The tetramethylbenzidine (TMB) method is a common chromogenic assay for hemoglobin.
    • Factors like albumin and bilirubin can interfere with TMB-based hemoglobin measurements.

    Purpose of the Study:

    • To investigate the influence of albumin on the hemoglobin-tetramethylbenzidine reaction.
    • To establish optimized conditions for accurate hemoglobin determination using TMB.
    • To adapt the TMB method for both automated centrifugal analyzers and manual assays.

    Main Methods:

    • Studied the hemoglobin-tetramethylbenzidine reaction kinetics using a centrifugal analyzer.
    • Varied albumin and bilirubin concentrations, incubation temperature, and reagent concentrations (TMB, hydrogen peroxide).

    Related Experiment Videos

  • Developed and validated procedures for automated and manual hemoglobin assays.

    • Main Results:

    • Albumin exhibits a biphasic effect, initially stimulating then inhibiting the reaction.
    • An intermediate incubation period was identified where the assay is independent of albumin and bilirubin.
    • Adjusting temperature, H2O2, and TMB concentrations altered the crossover time.
    • Automated assay demonstrated excellent accuracy (r=0.990 vs. direct method) and precision (CV < 3%).
    • Plasma recovery was good with citrate and heparin, but poorer with EDTA.

    Conclusions:

    • The TMB hemoglobin assay is significantly influenced by albumin, requiring careful optimization.
    • The identified crossover point allows for albumin- and bilirubin-independent hemoglobin measurement.
    • The developed method is accurate, precise, and adaptable to different analytical platforms.
    • Proper sample collection (avoiding EDTA) is important for reliable hemoglobin recovery.